Publicado

2017-04-01

Expresión de la proteína recombinante Cry 1Ac en cultivos de células de papa en suspensión: Establecimiento del cultivo y optimización de la producción de la biomasa y la proteína mediante la adición de nitrógeno

Expression of recombinant Cry 1Ac protein in potato plant cell suspension culture: Establishment of culture and optimization of biomass and protein production by nitrogen supply

Palabras clave:

Cultivo de células vegetales en suspensión, proteínas recombinantes, Solanum tuberosum (es)
Plant cell suspension culture, recombinant proteins, Solanum tuberosum (en)

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Autores/as

  • Carlos Julio Nova-López Universidad Nacional de Colombia, sede Medellín.
  • Jorge Mario Muñoz-Pérez Universidad Nacional de Colombia, sede Medelín
  • Luisa Fernanda Granger-Serrano Universidad Naciona de Colombia, sede Medellín.
  • Mario Eveilio Arias-Zabala Universidad Nacional de Colombia, sede Medellín.
  • Rafael Eduardo Arango-Isaza Universidad Nacional de Colombia, sede Medellín.
Los cultivos in vitro de células vegetales en suspensión se han propuesto como plataformas alternativas de expresión de proteínas recombinantes con aplicación terapéutica por las ventajas que ofrecen sobre los sistemas tradicionales de expresión en células bacterianas y de mamíferos. En este trabajo se determinó un protocolo para el establecimiento de suspensiones de papa (S. tuberosum) genéticamente modificadas con el gen de la proteína Cry 1Ac y se caracterizaron las cinéticas de producción de la biomasa y la proteína recombinante. Los entrenudos y el medio MS suplementado con 2.0 mg L-1 de 2,4-D, mostraron los mejores porcentajes de formación de callo. La tasa máxima de crecimiento específico calculada para las suspensiones fue 0.12 d-1, con una concentración máxima de biomasa de 1.41 g L-1 al final de la fase exponencial, la cual logró aumentarse hasta 3.94 g L-1 duplicando la concentración de NO3- y NH4+ en el medio de cultivo.
Plant cell suspension cultures have been proposed as alternative platforms for the expression of recombinant proteins with therapeutic application because of its potential advantages over traditional bacterial and mammalian cells-based platforms. In this work, a protocol for the establishment of potato suspensions (S. tuberosum) genetically modified with Cry 1Ac gene was obtained and the kinetics of biomass and recombinant protein production were characterized. Internodal explants and MS medium supplemented with 2.0 mg L-1 of 2,4- D showed the best response in terms of callus formation. The maximum specific growth rate for suspensions was 0.12 d-1, with a maximum biomass concentration of 1,41 g L-1 at the end of exponential phase. This biomass concentration was improved to 3.94 g L-1 bydoubling the concentration of NO3- and NH4+ in the culture medium.

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